不同种植年限黄连根系土壤细菌PCR―DGGE剖析.docVIP

不同种植年限黄连根系土壤细菌PCR―DGGE剖析 　　[摘要]该研究应用PCR-DGGE技术，分析了不同种植年限正常生长、发病的黄连Coptis chinensis的根际和非根际土壤的细菌种群多样性指数，并选取代表性DGGE条带进行克隆测序，构建细菌系统发育树状图。结果表明，正常土样与发病土样土壤细菌种群差异明显，且发病土样多样性指数（H）高于正常土样;代表性条带克隆测序结果表明，黄连种植土壤中未培养细菌为优势菌群，其余类群为食酸菌属、芽孢杆菌属、不动杆菌属、未培养克吕沃尔菌属以及未培养丛毛单胞菌，未发现已报道的植物病原细菌。发病土样中Clone band d的谱带亮度高于正常土样，该条带菌株在分类上属于食酸菌属。显然，黄连种植过程中，根际土壤中细菌种群发生了改变，其中食酸菌属的细菌在发病根际土壤增加，很可能与黄连病害发生有关。 　　[关键词]黄连;细菌种群;根际土壤;16S rDNA;PCR-DGGE 　　Bacteria community in different aged Coptis chinensis planting 　　soil revealed by PCR-DGGE analysis 　　TAN Yuan， CHEN Qiang*， LIU Han-jun， SONG San-duo， YU Xiu-mei， DONG Zhen-huan， TANG Xue， ZHONG Yu-zhou 　　（College of Resource Sciences and Technology， Sichuan Agricultural University， Wenjiang 611130， China） 　　[Abstracts]In order to reveal the cause of disease occurred in the process of Coptis chinensis growth， this paper studied the bacterial species diversity index of different aged rhizospheric and non-rhizospheric soil planting normal or sick C. chinensis by using PCR-DGGE technique. The representative DGGE bands were chosen to be cloned， and sequenced， the phylogeny were constructed. The results showed that the bacterial communities were very different between the normal and diseased soil samples of C. chinensis， and the diversity index （H） of diseased soil samples were higher than that of normal soil samples.Sequencing analysis of representative cloned DGGE bands showed that the unculturable bacteria were the dominant groups， and bacteria belonged to genus Bacillus， Acidovorax， Acinetobacter， uncultured Kluyvera， and uncultured Comamonas were also existing， but the reported plant pathogenic bacteria were not found in the C. chinensis planting soil. The density and brightness of clone band d in diseased soil samples was higher than that in normal soil sample， and sequencing analysis showed that it belonged to genus Acidovorax.Obviously， during the process of C. chinensis growth， the rhizospheric bacteria population changed， and the quantity of bacteri